Statistics for gene-specific qPCR primers designed for 147 organisms.

Organism	Source	Genome size (Mb)	Gene number	Best primer (level1)	Best primer (level2)	Best primer (level3)	Sum of best primers	Percentage of best primers	All primer (level1)	All primer  (level2)	All primer (level3)	Sum of all primers	Average primer paris
Apis mellifera	Ensembl	229.1	15320	9407	3340	127	12874	84.03%	65485	294355	26228	386068	25.20
Arabidopsis halleri	Phytozome	121	25008	15538	9108	95	24741	98.93%	57223	455371	39502	552096	22.08
Arabidopsis thaliana	TAIR	135.7	27416	15830	9995	170	25995	94.82%	58455	423831	38667	520953	19.00
Beta vulgaris ssp. vulgaris	Ensembl	517.4	26521	9849	11107	77	21033	79.31%	18966	349734	46870	415570	15.67
Boechera stricta	Phytozome	196.5	27416	15768	11065	168	27001	98.49%	31688	314284	33562	379534	13.84
Bombyx mori	Ensembl	431.7	14623	11698	2043	108	13849	94.71%	59017	102483	8615	170115	11.63
Bos taurus	Ensembl	2649.7	20791	12629	7192	149	19970	96.05%	30387	306893	69948	407228	19.59
Brachypodium distachyon	Ensembl	271.9	26552	17472	8179	105	25756	97.00%	39554	295413	55501	390468	14.71
Brachypodium stacei	Phytozome	234	29898	17418	11459	350	29227	97.76%	35780	365422	79488	480690	16.08
Brassica napus	Ensembl	738.4	101040	12679	70935	2959	86573	85.68%	14941	485323	129309	629573	6.23
Caenorhabditis briggsae	Ensembl	108.4	21996	12157	7303	201	19661	89.38%	30430	183965	16745	231140	10.51
Callithrix jacchus	Ensembl	2759.2	23323	12050	9407	323	21780	93.38%	26776	301349	61798	389923	16.72
Canis lupus familiaris	Ensembl	2392.7	20806	10300	8965	331	19596	94.18%	22182	379471	91133	492786	23.68
Capsella grandiflora	Phytozome	100.5	24805	15507	9022	106	24635	99.31%	33117	290202	28034	351353	14.16
Capsella rubella	Phytozome	134.8	26521	14722	10818	177	25717	96.97%	28789	307765	33028	369582	13.94
Carica papaya	Phytozome	135	27769	15469	8957	357	24783	89.25%	38727	166584	12310	217621	7.84
Cavia porcellus	Ensembl	2663.4	19065	12531	5814	194	18539	97.24%	35501	215727	41890	293118	15.37
Chenopodium quinoa	Phytozome	1385	44776	14261	26803	602	41666	93.05%	20465	333661	57716	411842	9.20
Chlamydomonas reinhardtii	Ensembl	120.4	14431	12528	995	185	13708	94.99%	80320	84681	40302	205303	14.23
Chlorocebus sabaeus	Ensembl	2762.7	19740	10289	8775	191	19255	97.54%	21105	396919	95744	513768	26.03
Choloepus hoffmanni	Ensembl	2060.4	14030	10565	2723	105	13393	95.46%	42045	131349	14309	187703	13.38
Cicer arietinum	LIS		28269	14322	10002	356	24680	87.30%	32811	214764	17401	264976	9.37
Ciona intestinalis	Ensembl	112.2	16698	13266	2930	45	16241	97.26%	43320	137851	8413	189584	11.35
Ciona savignyi	Ensembl	177	11832	9981	1274	40	11295	95.46%	44072	85821	4997	134890	11.40
Citrus clementina	Phytozome	301.4	24533	14796	8946	144	23886	97.36%	31306	292844	30475	354625	14.46
Citrus sinensis	Phytozome	319	25379	15144	9574	186	24904	98.13%	32537	272943	25844	331324	13.06
Cucumis melo	PLAZA		28812	15264	8856	438	24558	85.24%	37839	179555	13346	230740	8.01
Cucumis sativus	Phytozome	203	21503	14096	6721	92	20909	97.24%	33502	254571	21985	310058	14.42
Cyanidioschyzon merolae	Ensembl	16.7	5042	4686	122	18	4826	95.72%	46052	24333	4652	75037	14.88
Danaus plexippus	Ensembl	272.9	16317	12274	2976	166	15416	94.48%	54996	122903	10480	188379	11.54
Danio rerio	Ensembl	1464.4	26190	10638	14152	262	25052	95.65%	18638	437888	102715	559241	21.35
Dasypus novemcinctus	Ensembl		24211	10855	11827	429	23111	95.46%	22247	388558	94645	505450	20.88
Daucus carota	Phytozome	421	32113	16346	12997	301	29644	92.31%	33827	274856	26529	335212	10.44
Dipodomys ordii	Ensembl	1845	16808	12377	3832	136	16345	97.25%	42068	186725	31415	260208	15.48
Drosophila ananassae	Ensembl	213.9	15234	12136	2107	178	14421	94.66%	61232	133104	25384	219720	14.42
Drosophila erecta	Ensembl	145.1	15046	11895	2192	170	14257	94.76%	61276	132871	24278	218425	14.52
Drosophila grimshawi	Ensembl	186.1	14983	11052	2416	239	13707	91.48%	49704	124692	27318	201714	13.46
Drosophila melanogaster	Ensembl	142.6	14175	11459	2065	46	13570	95.73%	50178	214375	34696	299249	21.11
Drosophila mojavensis	Ensembl	180.2	14597	11737	1971	199	13907	95.27%	58027	125475	26949	210451	14.42
Echinops telfairi	Ensembl	2111.6	20399	14191	4841	273	19305	94.64%	51303	171450	29300	252053	12.36
Equus caballus	Ensembl	2428.8	24849	12979	9149	509	22637	91.10%	32765	261381	51208	345354	13.90
Erinaceus europaeus	Ensembl	2133.1	15992	11755	3416	145	15316	95.77%	43418	157832	23720	224970	14.07
Eucalyptus grandis	Phytozome	691	36349	16947	17654	402	35003	96.30%	29972	352847	57186	440005	12.11
Eutrema salsugineum	Phytozome	243.1	26351	15439	10141	163	25743	97.69%	32277	284269	30712	347258	13.18
Felis catus	Ensembl	2365.7	20035	11244	8133	213	19590	97.78%	27059	336208	71297	434564	21.69
Fragaria vesca	Phytozome	240	32831	15890	15444	455	31789	96.83%	30997	284396	34980	350373	10.67
Gadus morhua	Ensembl	608	20613	15811	3915	250	19976	96.91%	58758	168007	53345	280110	13.59
Gallus gallus	Ensembl	1285.6	18389	10430	6067	230	16727	90.96%	25013	269830	55208	350051	19.04
Gasterosteus aculeatus	Ensembl	446.6	20839	15540	4576	132	20248	97.16%	49034	247983	57440	354457	17.01
Glycine max	Ensembl	973.3	54174	14323	37418	555	52296	96.53%	18715	545054	125411	689180	12.72
Gossypium raimondii	Phytozome	761.4	37505	15988	20410	265	36663	97.75%	25208	476630	68390	570228	15.20
Heliconius melpomene	Ensembl	269.6	13637	10530	1808	35	12373	90.73%	53168	176147	11947	241262	17.69
Homo sapiens	Ensembl	3555	22357	11260	6057	315	17632	78.87%	54153	315861	55355	425369	19.03
Ictidomys tridecemlineatus	Ensembl	2311.1	19232	11324	7332	182	18838	97.95%	27562	268805	53954	350321	18.22
Ixodes scapularis	Ensembl	1388.5	20486	15125	3453	394	18972	92.61%	64201	99695	18195	182091	8.89
Kalanchoe fedtschenkoi	Phytozome	256	30964	15101	14979	237	30317	97.91%	26721	388097	57589	472407	15.26
Kalanchoe laxiflora	Phytozome	422	50461	10781	34886	0	45667	90.50%	13948	277988	65099	357035	7.08
Latimeria chalumnae	Ensembl	2183.6	19710	9899	9505	112	19516	99.02%	18796	374130	69596	462522	23.47
Leersia perrieri	Ensembl	266.7	29078	15543	11864	627	28034	96.41%	33060	334107	63729	430896	14.82
Lepisosteus oculatus	Ensembl	869.4	18383	10136	7939	110	18185	98.92%	21533	411851	92958	526342	28.63
Linum usitatissimum	Phytozome	318.3	43471	17709	22841	0	40550	93.28%	28729	263031	48870	340630	7.84
Lottia gigantea	Ensembl	298.9	23428	14472	5961	220	20653	88.16%	43995	208689	10977	263661	11.25
Lotus japonicus	LIS	472	29750	13415	11534	401	25350	85.21%	25653	171550	18004	215207	7.23
Loxodonta africana	Ensembl	3118.6	20601	13082	6773	247	20102	97.58%	36067	220997	40382	297446	14.44
Macaca mulatta	Ensembl	3246.4	21385	9769	9751	0	19520	91.28%	19462	408745	103302	531509	24.85
Manihot esculenta	Phytozome	495.48	33033	14763	15969	0	30732	93.03%	25084	390253	55113	470450	14.24
Marchantia polymorpha	Phytozome	225.8	19287	11995	6624	0	18619	96.54%	27375	341568	55249	424192	21.99
Medicago truncatula	Ensembl	412.8	50633	16632	27592	0	44224	87.34%	28184	455662	59918	543764	10.74
Megaselia scalaris	Ensembl	304.5	11482	8497	2183	0	10680	93.02%	32869	42840	1469	77178	6.72
Meleagris gallopavo	Ensembl	935.9	14247	9744	4195	0	13939	97.84%	26690	195393	29736	251819	17.68
Melitaea cinxia	Ensembl	361	16639	12676	3400	0	16076	96.62%	51780	163965	9775	225520	13.55
Microcebus murinus	Ensembl	2378	18311	10241	7107	0	17348	94.74%	23629	354780	84861	463270	25.30
Micromonas sp. RCC299	Phytozome		10103	9122	688	0	9810	97.10%	54579	59786	24874	139239	13.78
Micromonas pusilla CCMP1545	Phytozome	22	10660	8977	1106	0	10083	94.59%	44829	49409	27578	121816	11.43
Mimulus guttatus	Phytozome	312.7	28140	16442	10471	0	26913	95.64%	35095	311068	39220	385383	13.70
Mnemiopsis leidyi	Ensembl	150.4	17261	11244	4570	0	15814	91.62%	33076	214063	16096	263235	15.25
Monodelphis domestica	Ensembl	3501.7	22049	8730	11661	0	20391	92.48%	15471	433538	99450	548459	24.87
Mus musculus	Ensembl	3482.4	22233	12938	6876	266	20080	90.32%	58920	377031	66764	502715	22.61
Musa acuminata	Ensembl	390.6	36525	17453	13414	0	30867	84.51%	33691	307032	46610	387333	10.60
Myotis lucifugus	Ensembl	1966.4	21441	12711	7207	0	19918	92.90%	35548	216200	43642	295390	13.78
Nasonia vitripennis	Ensembl	239.6	19230	13310	4670	161	18141	94.34%	44821	197402	23654	265877	13.83
Nomascus leucogenys	Ensembl	2756.6	19767	10157	8993	0	19150	96.88%	19726	415355	100124	535205	27.08
Octopus bimaculoides	Ensembl	1984.3	33609	13690	14824	0	28514	84.84%	28892	374850	47000	450742	13.41
Oreochromis niloticus	Ensembl	815.7	21462	10303	10821	100	21224	98.89%	19246	442809	109863	571918	26.65
Ornithorhynchus anatinus	Ensembl	1917.7	22245	12283	8375	0	20658	92.87%	34663	259545	50956	345164	15.52
Oropetium thomaeum	Phytozome	250	28446	16377	8144	0	24521	86.20%	46441	195950	30760	273151	9.60
Oryctolagus cuniculus	Ensembl	2604	20294	10951	7899	0	18850	92.88%	27175	338181	84147	449503	22.15
Oryza barthii	Ensembl	308.3	34575	16911	14579	1131	32621	94.35%	36110	306590	60325	403025	11.66
Oryza brachyantha	Ensembl	260.8	32038	16101	11886	906	28893	90.18%	35766	275767	46436	357969	11.17
Oryza glaberrima	Ensembl	316.4	33164	17136	10678	779	28593	86.22%	41655	207787	38725	288167	8.69
Oryza sativa Indica	Ensembl	411.7	40745	19587	16928	1333	37848	92.89%	41665	277525	59239	378429	9.29
Oryza sativa Japonica	Ensembl	411.7	35679	17705	15443	525	33673	94.38%	33203	330383	66723	430309	12.06
Ostreococcus lucimarinus	Ensembl	13.2	7640	6402	514	0	6916	90.52%	41872	33044	8843	83759	10.96
Otolemur garnettii	Ensembl	2359.2	20809	12733	7237	0	19970	95.97%	32719	252789	48395	333903	16.05
Ovis aries	Ensembl	2534.3	21211	11594	8385	0	19979	94.19%	27763	306699	68970	403432	19.02
Pan troglodytes	Ensembl	2995.9	19331	10467	8158	140	18765	97.07%	21580	383034	87744	492358	25.47
Panicum hallii	Phytozome	554	37232	17605	16946	620	35171	94.46%	32316	415703	100124	548143	14.72
Papio anubis	Ensembl	2893.3	19930	10955	8151	205	19311	96.89%	25042	336992	79630	441664	22.16
Pediculus humanus	Ensembl	108.4	10787	8770	1623	36	10429	96.68%	42753	112811	7474	163038	15.11
Pelodiscus sinensis	Ensembl	2106.6	18286	8734	8714	169	17617	96.34%	16466	364094	77950	458510	25.07
Petromyzon marinus	Ensembl	647.4	10462	8865	878	96	9839	94.05%	52663	56149	12197	121009	11.57
Phaseolus vulgaris	Phytozome	537.2	27197	14061	12519	142	26722	98.25%	26181	357888	45705	429774	15.80
Physcomitrella patens	Ensembl	480	32273	16269	11841	479	28589	88.58%	36528	329491	41199	407218	12.62
Poecilia formosa	Ensembl	714.2	23675	10016	12726	204	22946	96.92%	17277	497083	140537	654897	27.66
Pongo abelii	Ensembl	3109.3	21447	10394	9527	290	20211	94.24%	21016	388152	94056	503224	23.46
Populus deltoides	Phytozome	446.8	44853	14290	23639	801	38730	86.35%	21962	409962	66749	498673	11.12
Populus trichocarpa	Ensembl	417.1	41377	16294	22762	550	39606	95.72%	25739	387039	58071	470849	11.38
Procavia capensis	Ensembl	2407.9	17177	13043	3639	126	16808	97.85%	45094	190713	32281	268088	15.61
Prunus persica	Ensembl	0	28108	14626	11140	266	26032	92.61%	30378	271538	30839	332755	11.84
Pteropus vampyrus	Ensembl	1839.4	18023	12809	4625	142	17576	97.52%	40358	213319	38507	292184	16.21
Rattus norvegicus	Ensembl	0	23845	11525	9661	344	21530	90.29%	24302	372254	85774	482330	20.23
Rhodnius prolixus	Ensembl	561.5	16506	11334	2407	72	13813	83.68%	47478	104595	4514	156587	9.49
Saccharomyces cerevisiae	Ensembl	12.2	6713	5425	491	17	5933	88.38%	37460	43733	1090	82283	12.26
Salix purpurea	Phytozome	392	37865	13530	19084	614	33228	87.75%	21625	366834	56595	445054	11.75
Sarcophilus harrisii	Ensembl	2931.6	18966	10144	8383	153	18680	98.49%	21312	321064	56273	398649	21.02
Sarcoptes scabiei	Ensembl	56.3	10657	7882	2193	74	10149	95.23%	31359	80596	5324	117279	11.00
Selaginella moellendorffii	Ensembl	212.6	34865	12405	13887	1091	27383	78.54%	22108	93772	22313	138193	3.96
Setaria italica	Ensembl	405.7	35471	19718	14039	447	34204	96.43%	40218	325029	67284	432531	12.19
Setaria viridis	Phytozome	394.9	35214	17878	16103	426	34407	97.71%	32605	435126	103427	571158	16.22
Sida fallax	Phytozome		26939	10874	15111	292	26277	97.54%	17231	416858	96507	530596	19.70
Solanum lycopersicum	Ensembl	737.6	33886	16016	14392	372	30780	90.83%	31284	308268	29330	368882	10.89
Solanum tuberosum	Ensembl	727.4	39353	17057	20080	491	37628	95.62%	30060	339655	36565	406280	10.32
Solenopsis invicta	Ensembl	354.7	14922	11010	3616	61	14687	98.43%	38114	245058	30416	313588	21.02
Sorghum bicolor	Ensembl	738.5	34496	18939	12381	651	31971	92.68%	40650	302510	62084	405244	11.75
Spirodela polyrhiza	Phytozome	158	19623	13992	4600	342	18934	96.49%	48262	130900	24065	203227	10.36
Strongylocentrotus purpuratus	Ensembl	816	40772	11644	14749	380	26773	65.67%	21494	324400	46931	392825	9.63
Strongyloides ratti	Ensembl	43.2	12449	9505	2133	44	11682	93.84%	35857	86419	4398	126674	10.18
Sus scrofa	Ensembl	3024.7	22198	11586	8347	299	20232	91.14%	27078	302217	67647	396942	17.88
Taeniopygia guttata	Ensembl	1222.9	17894	9939	5299	0	15238	85.16%	27315	174054	29571	230940	12.91
Takifugu rubripes	Ensembl	393.3	18685	13519	4381	0	17900	95.80%	42955	218639	44874	306468	16.40
Carlito syrichta	Ensembl	2768.6	14865	10888	3271	0	14159	95.25%	38545	158433	19628	216606	14.57
Tetranychus urticae	Ensembl	89.6	18020	11494	4153	0	15647	86.83%	37087	166861	10276	214224	11.89
Thellungiella parvula	PLAZA		25691	16021	9084	171	25276	98.38%	37789	232147	21643	291579	11.35
Tribolium castaneum	Ensembl	152.4	16741	11830	3597	166	15593	93.14%	44210	148745	14911	207866	12.42
Triticum urartu	Ensembl	3009	34977	18450	12405	755	31610	90.37%	43380	244927	40632	328939	9.40
Tupaia belangeri	Ensembl	2137.2	17784	12631	3874	216	16721	94.02%	47153	161593	24112	232858	13.09
Tursiops truncatus	Ensembl	2298.5	17475	12552	4379	141	17072	97.69%	39719	208509	39356	287584	16.46
Vicugna pacos	Ensembl	1923	12663	9933	2256	0	12189	96.26%	40016	129073	14310	183399	14.48
Vigna angularis	LIS		26857	14318	10509	275	25102	93.47%	31858	234398	21958	288214	10.73
Vigna radiata	LIS		22368	13506	8091	163	21760	97.28%	31935	214322	18557	264814	11.84
Vitis vinifera	Ensembl	486.2	29971	14333	12751	359	27443	91.57%	27510	282367	34360	344237	11.49
Volvox carteri	Phytozome		14247	12918	1056	65	14039	98.54%	79989	261557	87086	428632	30.09
Xenopus tropicalis	Ensembl	1358.3	18615	10951	6973	84	18008	96.74%	24073	286554	40985	351612	18.89
Xiphophorus maculatus	Ensembl	652.8	20407	10523	9509	141	20173	98.85%	20667	424328	102997	547992	26.85
Zea mays	Ensembl	2104.4	63480	17914	31891	1562	51367	80.92%	28688	441858	110699	581245	9.16
Zootermopsis nevadensis	Ensembl	464.3	14610	9805	3393	0	13198	90.34%	34933	129663	10001	174597	11.95
Zostera marina	Phytozome		20450	13576	5321	132	19029	93.05%	35968	215356	16615	267939	13.10
Total			3598514	1899842	1397989	36864	3334695	93.44%	5212329	39038395	6841061	51091785	15.09

Note: Ensembl (http://www.ensembl.org/index.html); LIS: Legume Information System (https://legumeinfo.org); Phytozome (http://www.phytozome.net);  PLAZA (http://bioinformatics.psb.ugent.be/plaza).